Abstract
| Original language | English |
|---|---|
| Journal | Methods in Molecular Biology |
| Volume | 873 |
| Pages (from-to) | 297-305 |
| Number of pages | 9 |
| ISSN | 1064-3745 |
| DOIs | |
| Publication status | Published - 2012 |
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Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis. / Harkness, Linda; Prokhorova, Tatyana A; Kassem, Moustapha; Blagoev, Blagoy.
In: Methods in Molecular Biology, Vol. 873, 2012, p. 297-305.Research output: Contribution to journal › Journal article › Research
TY - JOUR
T1 - Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis
AU - Harkness, Linda
AU - Prokhorova, Tatyana A
AU - Kassem, Moustapha
AU - Blagoev, Blagoy
PY - 2012
Y1 - 2012
N2 - The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.
AB - The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.
U2 - 10.1007/978-1-61779-794-1_20
DO - 10.1007/978-1-61779-794-1_20
M3 - Journal article
VL - 873
SP - 297
EP - 305
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -